Immunological Effect on Egg-laying Chickens by Combining Vaccinations Against Newcastle and Infectious Bursal Diseases
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摘要: 为探讨新城疫-传染性法氏囊病二联灭活疫苗（ND-IBD二联灭活苗）对不同日龄商品蛋鸡的免疫效果，选择360只1日龄商品蛋鸡，分别于1、7日龄接种ND-IBD二联灭活苗，对照组接种灭菌生理盐水，分别于21-56日龄，每隔7 d采血，分离血清，检测ND和IBD抗体水平，剖检取脾脏与法氏囊观察病变，荧光定量PCR检测免疫因子CD4+、CD8+、IL-6、IFN-βmRNA表达量，并进行攻毒保护试验。结果表明，1、7日龄商品蛋鸡接种ND-IBD二联灭活苗，其ND和IBD的抗体水平均有效提高，且在免疫后可以有效保护IBDV强毒的感染，但7日龄免疫效果更好。免疫因子的mRNA表达量检测结果表明，7日龄接种疫苗的免疫组免疫因子CD4+、CD8+、IL-6、IFN-β的mRNA表达量明显高于对照组1日龄。综上，ND-IBD二联灭活疫苗7日龄免疫商品蛋鸡免疫效果更好。Abstract: Immunological effect of combined inoculation of the inactivated vaccines for Newcastleand the infectious bursal diseases (ND-IBD) on egg-laying chickens was investigated. For the study, 360 one-day-old commercial chicks were randomly divided into 3 groups. The chicks in one group were immunized immediately with the two vaccines simultaneously, another when they were 7-day-old, and the third group injected with saline solution as control.Levels of ND-IBD antibodies in the serum ofthese chicks were determined, spleen biopsies for disease detection executed, and qPCR on mRNA expressions of CD4+, CD8+, IL-6 and IFN-β performed every 7 days during the period of 21 to 56 d. The results showed that all treated one-or 7-day-old birds had higher antibody levels than control, and could fence off IBD virus infection. The 7-day-old group showed greater immunity against the diseases, and significantly increased mRNA expressions on CD4+, CD8+, IL-6 and IFN-β over control. It was, therefore, concluded that, by combining vaccinations against ND-IBD on 7-day-old egg-laying chicks, a desirable immunological effect on the chickens could be achieved.
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Key words:
- newcastle disease /
- infectious bursal disease /
- inactivated vaccine /
- antibody detection
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图 1 不同处理IBD抗体水平
注：不同小写字母表示同一日龄不同处理间差异显著(P < 0.05)，下图同。
Figure 1. Levels of antibody against IBD in treatment groups
图 3 法氏囊组织中基因CD4、CD8、IL-6、IFN-β表达水平
注：A为商品蛋鸡法氏囊中CD4+基因表达量变化水平；B为商品蛋鸡法氏囊中CD8+基因表达量变化水平；C为商品蛋鸡法氏囊中IL-6基因表达量变化水平；D为商品蛋鸡法氏囊中IFN-β基因表达量变化水平。图 4同。
Figure 3. mRNA expressions of CD4+, CD8+, IL-6 and IFN-β in bursa tissue of chicks
图 4 各日龄鸡脾脏组织中基因CD4、CD8、IL-6、IFN-β表达量变化
Figure 4. mRNA expressions of CD4+, CD8+, IL-6 and IFN-β in spleen of chicks
表 1 引物序列
Table 1. Sequences of primers for RT qPCR
基因 引物序列(5′-3′) 扩增产物大小/bp CD4-F 5-GCAGTGGTGAGCTGTGTGTT-3 106 CD4-R 5-TCACCTCCTTTCCTGCAATC-3 CD8-F 5-CAGGGACAGAGGAACACGAT-3 125 CD8-R 5-CGAATCCAGGAGACACCATT-3 IL-6-F 5-ACCCAGAAATCCCTCCTCAC-3 126 IL-6-R 5-AAATAGCGAACAGCCCTCAC-3 IFN-β-F 5-GCTCTCCTGTTGTGCTTCTCCAC-3 120 IFN-β-R 5-CAATAGTCTCATTCCAGCCAGTGC-3 GAPDH-F 5-GCAGGAACATCCCGAAGAAGC-3 191 GAPDH-R 5-CCCGGAAGTGACCATGAGTAG-3 
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表 2 攻毒保护试验结果
Table 2. Survival rate of chicken artificially infected with IBDV
日龄/d 攻毒保护率/% A组(对照组) B组(1日龄免疫组) C组(7日龄免疫组) 21 25 70 80 28 5 80 90 35 0 100 100 42 0 100 100 49 0 100 100 56 0 100 100
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